The degree of effectiveness of mosquito nets against malaria in the Americas has remained uncertain. We carried out a case-control study of net use and mild malaria in the Amazonas state of Colombia. Two hundred ninety cases were enrolled via the Health Department services, and 977 community-based controls matched for age, sex, and place of residence. We found that a large proportion of the population (96% of controls) slept under nets. Nevertheless, we found a benefit of impregnated nets compared with no net use: adjusted odds ratio (OR) for mild malaria 0.44, 95% confidence interval (CI) 0.20–0.98. Nonimpregnated nets had a benefit that was only slightly smaller but not statistically significant (OR for mild malaria 0.54, 95% CI 0.25–1.18). Travel in the previous month had an odds ratio of 6.2 (95% CI 3.1–8.8) and a population attributable fraction of 13% compared with 11% for failure to use an impregnated net. We conclude that, in the Amazon region, promotion of mosquito net use and impregnation is justified, and that there is a need for measures to protect travelers from malaria.